Back to EveryPatent.com
United States Patent | 6,197,510 |
Vinayagamoorthy | March 6, 2001 |
The invention provides a method of sequencing a nucleic acid in a reaction mixture comprising first and second nucleic acid target sequences. The target sequences may be present on the same or different nucleic acid molecules. First and second labelled sequencing primers are provided that are hybridizable, respectively, to the first and second nucleic acid target sequences. The second sequencing primer being at least as long as the total length of the first sequencing primer plus the length of the first target sequence. Each of the sequencing primer extension products beginning with the second sequencing primer will be longer than the primer extension products in the pool beginning with the first sequencing primer. Once the sequencing reactions have been completed, the lengths of the primer extension products in the pool may be detected by known methods, such as by gel electrophoresis.
Inventors: | Vinayagamoorthy; Thuraiayah (Saskatoon, CA) |
Assignee: | Bio-Id Diagnostic Inc. (Saskatoon, CA) |
Appl. No.: | 165264 |
Filed: | October 1, 1998 |
Current U.S. Class: | 435/6; 435/5; 435/91.2 |
Intern'l Class: | C12P 019/34; C12Q 001/70 |
Field of Search: | 435/6,91.2,5 |
4683195 | Jul., 1987 | Mullis et al. | 435/6. |
4683202 | Jul., 1987 | Mullis et al. | 435/91. |
4965188 | Oct., 1990 | Mullis et al. | 435/6. |
5403707 | Apr., 1995 | Atwood et al. | 435/5. |
5552283 | Sep., 1996 | Diamandis et al. | 435/6. |
5582989 | Dec., 1996 | Caskey et al. | 435/6. |
5776737 | Jul., 1998 | Dunn | 435/91. |
Foreign Patent Documents | |||
WO 93/06243 | Apr., 1993 | WO. | |
WO 94/10315 | May., 1994 | WO. | |
WO 98/26095 | Jun., 1998 | WO. | |
WO 99/29900 | Jun., 1999 | WO. |
Li et al., "Direct electrophoretic detection of the allelic states of single DNA molecules in human sperm by using the polymerase chain reaction" Proc. Natl. Acad. Sci USA vol. 87, pp. 4580-4584, 1990. Rawn, "Biochemistry" pp. 666, 708, Carolina Biological Supply, North Carolina 1989. S. C. Roemer et al., Simultaneous Bi-Directional Cycle Sequencing, LI-COR Virtual Poster Session [on-line], September 1997. retrieved on Sep. 4, 1998, from Internet: <URL:http://www.licor.com/bio/Posters/GenSeq97/GenSeq97.htm>. D.L. Steffens et al., Multiplex Amplification of Human STR Loci using Infrared Fluorescence Detection, LI-COR Virtual Poster Session [on-line], Feburary, 1997. Visible Genetics Inc., Application Note, HPV Genotyping using the OpenGene Automated DNA Sequencing System, Jan. 16, 1998. Visible Genetics Inc., Application Note, HPV Genotyping using the OpenGene Automated DNA Sequencing System, Jan. 16, 1998. Visible Genetics Inc., Application Note, p53 Mutation Detection using the OpenGene Automated DNA Sequencing System, May 3, 1998. Jones, Leslie B. et al.: "Octamer-primed cycle sequencing using dye-terminator chemistry" Nucleic Acids Research, vol. 26, No. 11, 1998, pp. 2824-2426, XP002131455. Horton, Joseph H.: "Optimized Conditions for Cycle Sequencing of PCR Products" PCR Methods And Applications, vol. 3, 1994, pp. 359-360, XP002131456. White, Michael J. et al.: "Concatemer Chain Reaction: A Taq DNA Polymerase-Mediated Mechanism for Generating Long Tandemly Repetitive DNA Sequences" Analytical Biochemistry, US, Academic Press, San Diego, CA, vol. 199, No. 2, Dec. 1, 1991, pp. 184-190, XP000236491. Kretz, Keith et al.: "Cycle Sequencing" PCR Methods & Applications, US, Cold Spring Harbor Laboratory Press, vol. 3, No. 5, Apr. 1, 1994, pp. S107-S112, XP000606764. Sanger, F. et al.: "DNA Sequencing with Chain-Terminating Inhibitors" Proceedings of the National Academy of Sciences of the USA, US, New York, NY, vol. 74, No. 12, Dec. 1, 1977, pp. 5463-5467, XP000603873. |